RelB shRNA慢病毒感染小鼠骨髓树突状细胞的免疫学效应  

Effects of RelB shRNA lentivirus infection on immunological response of mouse bone marrow dendritic cells

作　　者：谢金敏[1,2] 包杰[3] 董瑞强[4] 杨明[4] 温浩[1] 

机构地区：[1]新疆医科大学第一附属医院临床医学博士后流动站,新疆维吾尔自治区乌鲁木齐市830011 [2]解放军第四七四医院普外科,新疆维吾尔自治区乌鲁木齐市830011 [3]南方医科大学南方医学院检验医学科,广东省广州市510515 [4]石河子大学医学院,新疆维吾尔自治区石河子市830011

出　　处：《中国组织工程研究与临床康复》2011年第53期9986-9990,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省自然科学基金资助项目(8451051501000580);课题名称:RelB shRNA慢病毒介导的树突细胞治疗RA的研究~~

摘　　要：背景:沉默树突状细胞发育成熟的关键基因核因子кB/RelB可构建新型致耐受树突状细胞?目的:探讨RelB shRNA转染小鼠骨髓树突细胞的生物免疫学功能的影响。方法:利用重组粒细胞-巨噬细胞集落刺激因子和重组白细胞介素4联合诱导小鼠骨髓树突状细胞;慢病毒载体将RelBshRNA转染致小鼠骨髓树突细胞后,分为未成熟树突状细胞、脂多糖刺激成熟、RelB基因沉默及脂多糖刺激RelB沉默的4组树突状细胞进行观察。结果:体外培养第6天脂多糖刺激组细胞表面可见大量细长的类似树枝的突起,其他3组细胞形态特征相似,呈圆形、皱缩状态,这3组细胞表面MHC-II类分子、CD86和CD40分子表达水平相当,但低于脂多糖刺激组;3组混合淋巴细胞反应中刺激T细胞增殖能力差异无显著性意义(P>0.05),但均较脂多糖刺激组显著降低(P<0.01);RelB基因沉默的树突状细胞分泌Th1细胞因子γ-干扰素和白细胞介素2的能力较低,分泌Th2白细胞介素10和白细胞介素4的能力较高(P<0.01),Th1/Th2细胞因子的比例与未成熟树突状细胞类似。说明RelBshRNA经慢病毒转染骨髓源性树突状细胞后,在细胞形态、表面分子表达、免疫学功能等方面均具有与未成熟树突状细胞相似的特点,且不能被脂多糖刺激成熟。BACKGROUND： Whether nuclear factor-kappa B/RelB, a key gene to maturation of silenced dendritic cells, can construct novel tolerogenic dendritic cells remains poorly understood？ OBJECTIVE： To investigate the effects of RelB shRNA lentivirus infection on immunological response of mouse bone marrow derived bone marrow dendritic cells. METHODS： Mouse bone marrow derived dendritic cells were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor （rmGM-CSF） and recombinant mouse interleukin-4 （rmIL-4）. RelB shRNA lentivirus was transfected into mouse bone marrow derived bone marrow dendritic cells and then divided into four groups for observation： immature dendritic cells, mature dendritic cells stimulated by lipopolysaccharide, RelB-silenced dendritic cells, and RelB-silenced dendritic cells stimulated by lipopolysaccharide. RESULTS AND CONCLUSION： After 6 days of in vitro culture, in the group of mature dendritic cells stimulated by lipopolysaccharide, a large number of slender branch-like processes on the cell surface; in the other three groups, cell morphology was similar, exhibiting a round and shrunk appearance, and MHC-II molecule, CD86 and CD40 expression levels were similar, but they were lower compared with the group of mature dendritic cells stimulated by lipopolysaccharide. There was no significant difference in ability to stimulate T cell stimulation among the latter three groups, but the ability in these three groups was significantly lower than that in the group of mature dendritic cells stimulated by lipopolysaccharide （P 〈 0.01）. RelB-silenced dendritic cells showed poor ability to secrete Th1 cytokineγ- interferon and IL-2 and strong ability to secrete Th2 cytokine IL-10 and IL-4 （P 〈 0.01）. The proportion of Th1/Th2 cytokines was similar between the group of RelB-silenced dendritic cells and group of immature dendritic cells. These findings suggest that after RelB shRNA lentivirus infection, bone marrow-derived dendritic cells exhibit simila

关 键 词：转录因子RelB 树突状细胞 RNA干扰 免疫耐受 未成熟 脂多糖 

分 类 号：R318[医药卫生—生物医学工程]